Neutron Reflectometry of clathrin-mediated endocytosis
Clathrin-mediated endocytosis is crucial for the internalisation of most eukaryotic cell surface proteins. Clathrin-coated vesicles (CCVs) assemble with their cargo at the plasma membrane then transport these to the early endosome inside the cell. A CCV consists of a clathrin scaffold coating a lipid vesicle, in which the cargo is bound, linked by adaptor proteins, like AP2, that are associated with effectors of CCV assembly, stability and disassembly. In vivo, AP2 solely interacts with one leaflet of the cellular membrane. Therefore, we have been able to analyse the first stages of CCV assembly by using cargo embedded in a lipid monolayer and Langmuir trough methods. Importantly, the addition of AP2, then of clathrin, results in distinct physical changes. Available structural information about CCV formation has however been indirect. EM has only been on mature CCVs, either lacking lipids and AP2 or with a mixture of proteins. Crystal structures of AP2 (without lipids) have also suggested that large structural changes occur on binding the cellular membrane. To clarify between these different scenarios, neutron reflectometry analysis of CCV formation will be very informative.
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ZACCAI Nathan; CICUTA Pietro; Armando Maestro and OWEN David J.. (2018). Neutron Reflectometry of clathrin-mediated endocytosis. Institut Laue-Langevin (ILL) doi:10.5291/ILL-DATA.8-02-804
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